• Title/Summary/Keyword: asparagine

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Purification and Characterization of an Antilisterial Bacteriocin Produced by Leuconostoc sp. W65

  • Oh, Se-Jong;Kim, Myung-Hee;Churey, John-J.;Worobo, Randy-W.
    • Journal of Microbiology and Biotechnology
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    • v.13 no.5
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    • pp.680-686
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    • 2003
  • This study was carried out to characterize the antilisterial substances produced by Leuconostoc sp. W65 and to evaluate the effects of pH, temperature, and time on inhibitory activity using response surface methodology. Leucocin W65, an antilisterial substance produced by Leuconostoc sp. W65, markedly inhibited the growth of Listeria monocytogenes, L. innocua, and L. ivanovii, whereas other pathogens including Gram-negative bacteria were not susceptible. The pH was the most effective factor with regard to bacteriocin activity, while temperature and time of heat treatment had no significant effect. Fifty percent of inhibitory activity remained after 22.8 min at pH 4.2 and $121^{\circ}C$. Leucocin W65 was purified by ammonium sulfate precipitation, hydrophobic interaction chromatography, and tricine-SDS-PAGE. Compositional analysis originally estimated the peptide to be 56 amino acids in length without asparagine, glutamine, and tryptophane. The sequence of partial N-terminal amino acid residues of purified bacteriocin was identified as follows: $NH_{2}-XGXAGVXPXGGQQPXVPLXYP$.

The Analysis of Canavanine Content in Leaves, Roots, and Xylem Exudate of Canavalia lineata (해녀콩(Canavalia lineata)의 잎, 뿌리 및 도관액에서 Canavanine의 함량분석)

  • 박경순
    • Journal of Plant Biology
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    • v.33 no.2
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    • pp.119-126
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    • 1990
  • The content of canavanine was measured and analyzed in leaves, roots and xylem exudate of Canavalia lneata. In non-nodulated plants, the cotyledons were removed after a week of sowing and the plants were grown for 3 weeks. The quantity of canavanine measured by canavanine specific-PCAF colorimetric assay was 9-10 $\mu$mol/g fresh wt. in leaves, 5-6 $\mu$mol/g fresh wt. in roots, and 0.3-0.5 $\mu$mol/ml in xylem exudate. When free amino acids of leaves, roots, and xylem exudate were analysed by HPLC, the relative proportion of asparagine plus glycine was the highest and canavanine was high secondarily. And the relative proportion of canavanine among total free amino acids was 30-35% in leaves and roots, and 12-13% in xylem exudate. In non-nodulated plants grown for 8 weeks, the canavanine content of each part was similar to that of 3-week-old plants. By the formation of nodules, the canavanine content of leaves, roots, xylem exudate, and nodules decreased apparently. In xylem exduate, the nitrogenous compounds were also analyzed. The relative contents of NO3-, free amino acids, and ureides(allantoin and allantoic acid) were 60-80%, 20-30%, and 5%, respectively. From these results, it can be assumed that canavanine is synthesized in the root of plant and nodulation affects the canavanine content. It is obvious that canavanine is considered one of the reduced-N forms transported via xylem.

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Microarray analysis of hypoxia-induced changes in gene expression in BV-2 microglial cells (BV-2 microglia 세포주에서 저산소증의 유전자 발현에 대한 마이크로어레이 분석)

  • Kim, Bum-Shik;Seo, Jung-chul
    • Journal of Acupuncture Research
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    • v.20 no.4
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    • pp.85-92
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    • 2003
  • 목적 : 허혈시 발생되는 저산소중 상태에서는 세포독성을 유발한다고 알려져 있으나 정확한 기전은 아직 규명되지 않았다. 본 연구에서는 뇌허혈로 인한 세포독성의 기전을 유전자 발현을 통하여 살펴보고자 하였다. 방법 : 본 실험에서는 BV-2 microglia 세포주에 12시간 동안의 저산소 상태에서의 유전자 발현을 분석하기 위하여 마이크로에레이를 시행하였다. 결과 : 저산소 상태에서는 정상에 비하여 cathepsin F, growth factor independent 1, calcitonin/calcitonin-related poly, leucine-rich repeat LGI family membrane, dublecortin, cyclohydrolase 1, Ia-associated invariant chain, carbohydrate kinase-like과 erythrocyte protein band 4.1-like 3 등의 유전자 발현이 3배 이상 증가하였다. 한편 neuronal guanine nucleotide exchange factor, Bcl-2-related ovarian killer protein, chemokine (C-X-C motif) ligand 5, RNA binding motif protein 3, interleukin 2 receptor, alpha chain, crystallin zeta, cytochrome P450 subfamily IV B, asparagine synthetase과 moesin 등의 유전자 발현은 0.2배 이하로 감소하였다. 결론 : 이상의 결과는 저산소중에 관여하는 유전자 및 저산소중과 관련된 뇌경색 등의 질환의 기전을 밝히는데 기초적 자료로 이용될 수 있을 것이다.

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Protein Engineering of Deoxynucleoside Kinase from Lactobacillus acidophilus: Effect of Site-Directed Mutagenesis on Microbial Growth

  • Park, Inshik;Kim, Eun-Ae;Bang, Keuk-Seung;Kim, Seok-Hwan;Kim, Gi-Nahm;Lee, Min-Kyung;Kil, Ji-Oeun
    • Preventive Nutrition and Food Science
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    • v.6 no.1
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    • pp.79-81
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    • 2001
  • Deoxynucleoside kinases exist as heterodimeric pairs specific for deoxyadenosine/deoxyguanosine kinase (dAK/dGK) and deoxyadenosine/deoxycytidine kinase (dAK/dCK). The aspartic acid-84 in dGK was mutated to alanine, asparagine and glutamic acid by site-directed mutagenesis. The mutation resulted in a drastic decease in dGK activity compared to the unmodified cloned enzyme while it increased production of dAK activity. The mutated dak/dgk genes, which synthesize tandem deoxyadenosine/deoxyguanosine kinase, were inserted back to the Lactobacillus acidophilus and Lactococcus lactis by electroporation to determine the effect of site-directed mutation of he enzymes on the microbial growth. However, no significant change was observed in cell growth and lactic acid production between wild type and mutant lactic acid bacteria.

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The Content Analysis of Amino Acids Including GABA of Chlorella protothecoides under Mixtrophic Culture (혼합영양 배양에서 Chlorella protothecoides의 GABA를 포함한 아미노산 함량 분석)

  • Jeong, Yu Jeong;Kim, Seong Hak;Min, Hee Gyung;Kim, Sung Chun
    • Journal of Marine Bioscience and Biotechnology
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    • v.10 no.1
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    • pp.18-25
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    • 2018
  • Chlorella is quantitatively and qualitatively high in protein with balanced essential amino acid profiles, vitamins and minerals. ${\gamma}-Aminobutyric$ acid (GABA) is broadly distributed in nature and fulfills multi-physiological functions including effect such as a health-promoting functional compound. To improve the GABA production, Chlorella protothecoides were grown through the modified mixtrophic culture medium containing 2L of sterilized bristol medium with 0.01% urea and 4.0% glucose in a 5L fermenter. The results showed that nineteen kinds of amino acid including GABA at C. protothecoides sample were analyzed using high performance liquid chromatography (HPLC). Glutamic acid in total concentration (%) of amino acid is the most abundant amino acid (33.10%), followed by alanine (20.48%) and GABA (17.48%). Three amino acids including GABA were responsible for more than 70% total concentration in C. protothecoides sample including eight essential and nine non-essential amino acids: aspartic acid, asparagine, serine, glutamine, histidine, glycine, threonine, arginine, tyrosine, valine, methionine, tryptophan, phenylalanine, isoleucine, leucine, lysine. As a result of this experiment, it is expected that Chlorella will be developed to a critical product having high value as, GABA, functional food materials.

Synthesis of Insulin A (1-21) Chain and Their Assembly on a Polymer-Bound ${\alpha}$-Methylphenacylester Linkage (2-브로모프로피오닐화된 수지를 이용한 인슐린 A (1-21) 사슬의 합성)

  • Soon Uoong Koock;Nam-Joo Hong
    • Journal of the Korean Chemical Society
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    • v.31 no.5
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    • pp.457-463
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    • 1987
  • The total synthesis of insulin A chain (1-21) with properly protected sulfhdryl groups of three cysteins for the correct intra and inter disulfide bond formation has been accomplished on 2-bromopropionylated 2% DVB-styreneresin support employing manually operated rotary vessel. The sulfhydryl groups of cysteins were protected with acetamidomethyl, benzyl, and benzhydryl respectively. Glutamine and asparagine were attached to the peptide chain by active ester coupling, all other amino acids were coupled with DCC/HOBT. The synthesized peptide was purified by DEAE Sephadex A-25 and gel filtration Sephadex LH-20. The final product was found to be homogeneous by HPLC, electrophoresis, and amino acid analysis. The overall yield of the pure isolated peptide was 6%.

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Studies on the Physiological Chemistry of Spring Habits in Naked Barley II. Variration of Free Amino Acids during the Germination of Naked Barley with Different Spring Habits (과맥의 파성에 대한 생리화학적 연구 II. 파성심도에 따른 종자발아과정에 있어서의 유이아미노산의 소장)

  • 최선영
    • Journal of Plant Biology
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    • v.20 no.2
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    • pp.83-89
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    • 1977
  • Changes in the alcohol-soluble free amino acids during germination of a spring grain, Wanju and two winter grains, Sedohadaka and Nonsankwa No.1-6 which are differing in their degree of spring (winter) habits, were investigated by thin layer chromatography. The results obtained are as follows. 1. Throughout the germination period, 25 ninhydrin positive components; 22 amino acids including two amides and 3 unknown spots were detected. It is interesting to note that identification of histidine was confined to Wanju and Sedohadaka but Nonsankwa No.1-6, which has the lower degree of spring habit. 2. Except the quiescent seeds, the major components were generally composed of the acidic and neutral amino acids together with glutamine and asparagine. 3. Proline was contained in higher quantity except from the stage of quiescent seeds, but the outstanding difference among the varieties was not recognized. Whether this component is related to the mechanism of spring habit in barley or not is a problem to be studied more. 4. In all the varieties, most of the changes in amino acid levels during germination were usually in the same direction and of the same pattern under the temperature controlled not to be varnalized. In view of the results above, the changes in the amino acid levels seem to be more affected by the changes of growing circumstances and the subsequent metabolic activities of certain enzymes than by the characteristics of varieties themselves.

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Plant Regeneration from Embryogenic Suspension Cultures of Soybean (Glycine max L. Merrill)

  • Jang, Gi-Won;Park, Ro-Dong;Kim, Kwang-Soo
    • Journal of Plant Biotechnology
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    • v.3 no.2
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    • pp.101-106
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    • 2001
  • In order to establish efficient plant regeneration from embryogenic suspension cultures of soybean, Glycine max L, we examined the effects of auxin type and concentration, cytokinin type and concentration, and amino acid type and concentration on the growth of embryogenic clumps from induced callus, and the effect of desiccation of mature somatic embryos obtained from these clumps on the frequency of somatic embryo germination. Embryogenic callus was induced from the edge of the cotyledons cultured on MS medium containing 6% sucrose, 40 mg/L 2,4-D, 0.2% gelrite and pH 5.7. The growth of embryogenic clumps was best in early staged, embryogenic callus that was placed in suspension culture of MS medium containing 5 mg/L 2,4-D and 0.5 mg/L asparagine. Single somatic embryos were isolated from the clumps and plated on the same medium for maturation. When the mature single somatic embryos were desiccated for 96 h, somatic embryo germination came up to approximately 90%. The plantlets germinated after embryos desiccation for 2 weeks were transfered to MS medium containing 3% sucrose,0.2% gelrite and pH 5.7.

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Compositions of Extractive Nitrogenous Constituents and Their Monthly Variation for Fresh Capsosiphon fulvescens

  • Jung, Kyoo-Jin;Park, Jung-Nim
    • Preventive Nutrition and Food Science
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    • v.15 no.2
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    • pp.120-129
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    • 2010
  • To elucidate the composition of extractive nitrogenous components in the fresh Capsosiphons fulvescens cultured off the southern coast of Korea, and to determine the monthly variation of these nitrogenous components, extract samples collected monthly from December to March at Jangheung-gun, Jeonnam Province were analyzed for total nitrogen, free and combined amino acids, ATP and related compounds, betaines, trimethylamine oxide (TMAO) and trimethylamine (TMA). The content of extractive nitrogen was 1,090~1,233 mg/100 g on dry basis. The number of 21~25 ninhydrin-positive substances was detected in the analysis of free amino acids, and their total amount was 3,710~4,788 mg/100 g on dry basis. Among them, free proline, asparagine, glutamic acid, alanine, taurine and glutamine were found to be abundant. The combined amino acids amounted to 1,573~2,121 mg/100 g in total and the total amount of ATP and related compound was 33.8~84.0 mg/100 g ($1.06{\sim}2.46\;{\mu}mol/g$) on dry basis. Betaine, glycinebetaine, $\beta$-alaninebetaine, $\gamma$-butyrobetaine, homarine and trigonelline were detected in most of samples. Levels of free and combined amino acids, ATP and related compounds fluctuated from sample to sample, with their contents higher in December and January and lower in March.

Analysis for nucleotide sequence of the membrane protein gene of porcine epidemic diarrhea virus Chinju99

  • Baquilod, Greta Salvae V.;Yeo, Sang-Geon
    • Korean Journal of Veterinary Research
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    • v.46 no.4
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    • pp.355-361
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    • 2006
  • Porcine epidemic diarrhea virus (PEDV) strain Chinju99, which was previously isolated from piglets suffering from severe diarrhea was used to characterize the membrane (M) protein gene to establish the molecular information, and the results will be useful in elucidating concepts related to molecular pathogenesis and antigenic structures of PEDV isolates. The Chinju99 M gene generated by reverse transcription and polymerase chain reaction (RT-PCR) consisted of 681 bases containing 22.3% adenine, 22.3% cytosine, 23.1% guanine and 32.3% thymine nucleotides, and the GC content was 45.4%. It had some nucleotide mismatches from M gene of other PEDV strains, such as CV777, Br1/87, KPEDV-9, JMe2, JS2004-2 and LJB-03 with 97-99% nucleotide sequence homology to these strains. Also, it encoded a protein of 226 amino acids, which had some mismatches from those of CV777, Br1/87, KPEDV-9, JMe2, JS20004-2 and LJB-03, as the amino acid sequence homology showed a 97-98% to these strains. The Chinju99 had a very close relationship to the Japanese strain JMe2 for the nucleotide and amino acid sequences of the M gene. The amino acids predicted from Chinju99 M gene consisted of mostly hydrophobic residues and contained three potential sites for asparagine (N)-linked glycosylation, two serine (S)-linked phosphorylation sites by protein kinase C, and two S- or threonine (T)-linked phosphorylation sites by casein kinase II.